Journal: Scientific Reports

Article Title: Insulin signaling and glucose metabolism in different hepatoma cell lines deviate from hepatocyte physiology toward a convergent aberrant phenotype

doi: 10.1038/s41598-020-68721-9

Figure Lengend Snippet: The HepG2 human hepatoma cell line displays aberrant insulin signaling compared to murine primary hepatocytes. ( a ) Immunoblot analysis of insulin signaling in primary mouse hepatocytes infected with a control adenovirus expressing the GFP or a virus expressing the negative dominant RAS mutant H-RAS17N and stimulated with 10 nM insulin for 8 min. ( b ) Quantification of blots in ( a ). ( c ) Immunoblot analysis of insulin signaling and insulin receptor phosphorylation performed as in ( a ) but using the HepG2 cell line. ( d ) Quantification of blots in ( c ). n = 3 biological replicates (mice) for primary hepatocytes and 3 independent experiments for HepG2. Data are represented as mean ± SEM.

Article Snippet: Thereby, we have examined the information available on the HepG2 cell line at Broad Institute Cancer Cell Line encyclopedia (CCLE), where we have found 386 entries for mutations in the HepG2 genome.

Techniques: Western Blot, Infection, Control, Expressing, Virus, Mutagenesis, Phospho-proteomics

Journal: Scientific Reports

Article Title: Insulin signaling and glucose metabolism in different hepatoma cell lines deviate from hepatocyte physiology toward a convergent aberrant phenotype

doi: 10.1038/s41598-020-68721-9

Figure Lengend Snippet: Hepatoma cell lines display severely impaired gluconeogenic gene expression and blunted glucose production. ( a ) qPCR analysis of mRNA levels of glucose 6 phosphatase (G6P) and phosphoenolpyruvate carboxykinase (PEPCK) in primary mouse hepatocytes exposed for 6 h to 100 µM of dbcAMP, or 10 nM insulin, or 100 µM dbcAMP and 10 nM insulin. ( b ) qPCR analysis of mRNA levels of G6P and PEPCK in Hepa 1–6 cells treated as in ( a ). ( c ) qPCR analysis of mRNA levels of G6P and PEPCK in McARH7777 cell line treated as in ( a ). ( s ) qPCR analysis of mRNA levels of G6P and PEPCK in HepG2 cells treated as in ( a ). ( e ) Direct comparison of basal and cAMP-induced G6P and PEPCK mRNA levels in primary mouse hepatocytes and mouse Hepa 1–6. ( f ) Immunoblot analysis of insulin-driven AKT Thr 308 phosphorylation, AKT Ser 473 phosphorylation and GSK3β phosphorylation in primary hepatocytes and Hepa 1–6 cells stimulated for 8 min with 10 nM insulin. ( g ) Quantification of blots in ( f ). ( h ) Relative glucose production in primary hepatocytes, Hepa 1–6, McARH7777, and HepG2 stimulated with 100 µM dbcAMP in presence or not of 10 nM insulin for 6 h. n = 8 for ( a ); n = 5 for ( b ); n = 8 for ( c ); n = 7 for ( d ); and ( e ) n = 8 for heaptocytes and n = 5 for Hepa 1–6 n = 3 for ( f , g ) n = 3; for ( h ) n = 5 hepatocytes and n = 4 for each hepatoma cell line. n indicates biological replicates for primary hepatocytes, and independent experiments for cell lines. Data are represented as mean ± SEM.

Article Snippet: Thereby, we have examined the information available on the HepG2 cell line at Broad Institute Cancer Cell Line encyclopedia (CCLE), where we have found 386 entries for mutations in the HepG2 genome.

Techniques: Gene Expression, Comparison, Western Blot, Phospho-proteomics

Journal: Scientific Reports

Article Title: Insulin signaling and glucose metabolism in different hepatoma cell lines deviate from hepatocyte physiology toward a convergent aberrant phenotype

doi: 10.1038/s41598-020-68721-9

Figure Lengend Snippet: Hepatoma cell lines display reduced fatty acid synthase expression and a distinct protein electrophoresis pattern. ( a ) Immunoblot analysis of FAS on primary hepatocytes, Hepa 1–6, McARH7777, and HepG2. ( b ) Quantification of blots in ( a ). ( c ) Coomassie staining of a protein electrophoresis of extracts from human primary hepatocytes, murine primary hepatocytes, Hepa 1–6, McARH7777, and HepG2. n = 9 for a and b; n = 2–3 for c. n indicates biological replicates for primary hepatocytes, technical replicates for human hepatocytes, and independent experiments for cell lines. Data are represented as mean ± SEM.

Article Snippet: Thereby, we have examined the information available on the HepG2 cell line at Broad Institute Cancer Cell Line encyclopedia (CCLE), where we have found 386 entries for mutations in the HepG2 genome.

Techniques: Expressing, Protein Electrophoresis, Western Blot, Staining